A microfluidic device containing membrane-immobilized antibodies for successively capturing cytosolic enzymes
A microfluidic device containing membrane-immobilized antibodies for successively capturing cytosolic enzymes
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一种含有膜固定抗体的微流体装置，用于连续捕获胞质酶
DOI：
10.1016/j.talanta.2014.03.001
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发表时间：
2014
期刊：
影响因子：
6.1
通讯作者：
Ai. Hashimoto
中科院分区：
文献类型：
--
作者：
Youji Shimazaki;Ai. Hashimoto
制备了含有膜固定的抗酯酶（ES）抗体和抗乳酸脱氢酶（LDH）抗体的微流控装置。通过将已经通过非变性二维电泳分离的抗体转移到聚偏二氟乙烯膜上，然后将其染色并切成小片（16 mm 2）来制备膜。在该微流体装置中，>0.014单位mL-1的纯化的猪羧酸酯酶被膜固定的抗ES抗体特异性捕获，并且>147单位mL-1的纯化的猪LDH被膜固定的抗LDH抗体特异性捕获。此外，在猪肝细胞质中的微量等分试样的ES和LDH连续捕获膜固定化抗体的装置，并通过荧光分光光度法定量分析的酶的活性。结果表明，含有膜固定化抗体的微流控装置可用于研究几种类型的完整酶的活性。
A microfluidic device containing membrane-immobilized anti-esterase (ES) antibodies and anti-lactate dehydrogenase (LDH) antibodies was prepared. The membrane was prepared by transferring antibodies that had been separated by non-denaturing two-dimensional electrophoresis to a polyvinylidene difluoride membrane, which was then stained and cut into small pieces (16 mm2). In this microfluidic device, >0.014 Unit mL−1of the purified porcine carboxylesterase was specifically captured by membrane-immobilized anti- ES antibodies and >147 Unit mL−1of purified porcine LDH was specifically captured by membrane-immobilized anti-LDH antibodies. Furthermore, ES and LDH in micro-scale aliquots of porcine liver cytosol were successively captured by membrane-immobilized antibodies in the device, and the enzyme activities were quantitatively analyzed by spectrofluorometry. The results indicate that the microfluidic device containing membrane-immobilized antibodies can be used to investigate the activities of several types of intact enzymes.